Characterization of Pattern Recognition Receptor Expression and Functionality in Liver Primary Cells and Derived Cell Lines
Characterization of Pattern Recognition Receptor Expression and Functionality in Liver Primary Cells and Derived Cell Lines
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DOI：
10.1159/000489966
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发表时间：
2018-01-01
影响因子：
5.3
通讯作者：
Lucifora, Julie
中科院分区：
文献类型：
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作者：
Faure-Dupuy, Suzanne;Vegna, Serena;Lucifora, Julie
不同的肝细胞类型具有免疫学特性，包括细胞固有的先天免疫功能，这些功能对于最初控制病原体感染很重要。然而，主要人类肝细胞中先天免疫信号传导途径的表达和功能的完整景观仍然缺失。为了比较表征这些途径，我们从人肝切除中纯化了原代人肝细胞、肝星状细胞、肝窦内皮细胞（LSEC）和枯否细胞（KC）。我们评估了主要先天免疫传感器的mRNA和蛋白质表达水平，以及纯化细胞中的检查点抑制剂配体，并发现Toll样受体（TLR），RIG-I样受体，以及几种DNA胞质传感器在肝脏微环境中表达。在测试的细胞中，KC显示出在用PRR配体刺激时最广泛的活性，强调了它们在先天免疫感测肝脏微环境中的主要作用。通过KC永生化，我们产生了与常规用于研究单核细胞/巨噬细胞功能的THP 1细胞相比保留更高先天免疫功能的细胞系。我们的发现和KC系的建立将有助于了解TLR激动剂或检查点抑制剂抗病毒作用背后的免疫机制，这些药物目前正处于临床前或临床开发阶段。(C)2018 S. Karger AG，巴塞尔
Different liver cell types are endowed with immunological properties, including cell-intrinsic innate immune functions that are important to initially control pathogen infections. However, a full landscape of expression and functionality of the innate immune signaling pathways in the major human liver cells is still missing. In order to comparatively characterize these pathways, we purified primary human hepatocytes, hepatic stellate cells, liver sinusoidal endothelial cells (LSEC), and Kupffer cells (KC) from human liver resections. We assessed mRNA and protein expression level of the major innate immune sensors, as well as checkpoint-inhibitor ligands in the purified cells, and found Toll-like receptors (TLR), RIG-I-like receptors, as well as several DNA cytosolic sensors to be expressed in the liver microenvironment. Amongst the cells tested, KC were shown to be most broadly active upon stimulation with PRR ligands emphasizing their predominant role in innate immune sensing the liver microenvironment. By KC immortalization, we generated a cell line that retained higher innate immune functionality as compared to THP1 cells, which are routinely used to study monocyte/macrophages functions. Our findings and the establishment of the KC line will help to understand immune mechanisms behind antiviral effects of TLR agonists or checkpoint inhibitors, which are in current preclinical or clinical development. (C) 2018 S. Karger AG, Basel